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Two sulphur-oxidizing, chemolithoautotrophic aerobes were isolated from the chemocline of an anchialine sinkhole located within the Weeki Wachee River of Florida. Gram-stain-negative cells of both strains were motile, chemotactic rods. Phylogenetic analysis of the 16S rRNA gene and predicted amino acid sequences of ribosomal proteins, average nucleotide identities, and alignment fractions suggest the strains HH1 T and HH3 T represent novel species belonging to the genus Thiomicrorhabdus . The genome G+C fraction of HH1 T is 47.8 mol% with a genome length of 2.61 Mb, whereas HH3 T has a G+C fraction of 52.4 mol% and 2.49 Mb genome length. Major fatty acids of the two strains included C 16 : 1 , C 18 : 1 and C 16 : 0 , with the addition of C 10:0 3-OH in HH1 T and C 12 : 0 in HH3 T . Chemolithoautotrophic growth of both strains was supported by elemental sulphur, sulphide, tetrathionate, and thiosulphate, and HH1 T was also able to use molecular hydrogen. Neither strain was capable of heterotrophic growth or use of nitrate as a terminal electron acceptor. Strain HH1 T grew from pH 6.5 to 8.5, with an optimum of pH 7.4, whereas strain HH3 T grew from pH 6 to 8 with an optimum of pH 7.5. Growth was observed between 15–35 °C with optima of 32.8 °C for HH1 T and 32 °C for HH3 T . HH1 T grew in media with [NaCl] 80–689 mM, with an optimum of 400 mM, while HH3 T grew at 80–517 mM, with an optimum of 80 mM. The name Thiomicrorhabdus heinhorstiae sp. nov. is proposed, and the type strain is HH1 T (=DSM 111584 T =ATCC TSD-240 T ). The name Thiomicrorhabdus cannonii sp. nov is proposed, and the type strain is HH3 T (=DSM 111593 T =ATCC TSD-241 T ).
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Comparative genome analysis of test algal strain NIVA-CHL1 (Raphidocelis subcapitata) maintained in microalgal culture collections worldwide
Raphidocelis subcapitata is one of the most frequently used species for algal growth inhibition tests. Accordingly, many microalgal culture collections worldwide maintain R . subcapitata for distribution to users. All R . subcapitata strains maintained in these collections are derived from the same cultured strain, NIVA-CHL1. However, considering that 61 years have passed since this strain was isolated, we suspected that NIVA-CHL1 in culture collections might have acquired various mutations. In this study, we compared the genome sequences among NIVA-CHL1 from 8 microalgal culture collections and one laboratory in Japan to evaluate the presence of mutations. We found single-nucleotide polymorphisms or indels at 19,576 to 28,212 sites per strain in comparison with the genome sequence of R . subcapitata NIES-35, maintained at the National Institute for Environmental Studies, Tsukuba, Japan. These mutations were detected not only in non-coding but also in coding regions; some of the latter mutations may affect protein function. In growth inhibition test with 3,5-dichlorophenol, EC50 values varied 2.6-fold among the 9 strains. In the ATCC 22662–2 and CCAP 278/4 strains, we also detected a mutation in the gene encoding small-conductance mechanosensitive ion channel, which may lead to protein truncation and loss of function. Growth inhibition test with sodium chloride suggested that osmotic regulation has changed in ATCC 22662–2 and CCAP 278/4 in comparison with NIES-35.
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- Award ID(s):
- 1755220
- PAR ID:
- 10298899
- Editor(s):
- Cao, Yi
- Date Published:
- Journal Name:
- PLOS ONE
- Volume:
- 15
- Issue:
- 11
- ISSN:
- 1932-6203
- Page Range / eLocation ID:
- e0241889
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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Accepted Manuscript1.0
- Journal Article:
- https://doi.org/10.1371/journal.pone.0241889
